Process for producing beta-carotene

ABSTRACT

A PROCESS FOR PRODUCING INCREASED TITERS OF B-CAROTENE WHICH COMPRISES FERMENTING BLAKESLEA TRISPORA IN AN AQUEOUS FERMENTATION MEDIUM CONTAINING AT LEAST 7 1/2% CITRUS MOLASSES.

United States Patent 3,752,740 PROCESS FOR PRODUCING ,B-CAROTENE Oldrich K. Sebek, Kalamazoo, Mich., assignor to The Upjohn Company, Kalamazoo, Mich. No Drawing. Filed Jan. 22, 1964, Ser. No. 339,331 Int. Cl. C12d /00 U.S. Cl. 195-28 R 3 Claims ABSTRACT OF THE DISCLOSURE A process for producing increased titers of fi-carotene which comprises fermenting Blakeslea trispora in an aqueous fermentation medium containing at least 7 /2% citrus molasses.

This invention relates to an improved method for the production of fi-carotene by the fermentation of Blakeslea trisopora in an aqueous fermentation medium. More particularly, this invention relates to the discovery that high titers of i-carotene can be obtained by submerged aerobic fermentation of Blakeslea trispora using a novel fermentation medium containing at least T /2% citrus molasses.

B-Carotene has been obtained in media known in the art by means of Blakeslea trispora. For example, U.S. Pat. 3,079,380 discloses a medium for B-carotene. Citrus molasses is disclosed as being a valuable nutrient when used in levels below 5% While engaged in a study of synthesizing p-carotene by Blakeslea trispora, I discovered a novel fermentation medium which contains at least 7 /2% citrus molasses. My novel fi-carotene fermentation medium gives 40% and higher titers of p-carotene than any prior art medium known and tested by me.

Citrus molasses is a by-product of the citrus industry and contains approximately 72% solids in its marketable form. It is obtained from whole citrus peel by treating the peel with alkali, such as lime, and then pressing out the aqueous molasses from the peel.

My invention is a novel fl-carotene fermentation medium containing at least 7 /2% citrus molasses and, also, the use of at least 7 /2% citrus molasses in prior art [3- carotene fermentation media. For example, the fermentation medium and conditions in U.S. Pat. 3,079,380 can be used where the citrus molasses is substituted for the aqueous extract of citrus meal, as in Example 2.

The titer of fl-carotene in the fermentation broth can be determined by assay procedures disclosed in U.S. Pat. 3,079,380, Example 1 and U.S. Pat. 3,025,221, Example 1.

The following examples are illustrative of the medium and process of the present invention, but are not to be construed as limiting. Unless otherwise specified, parts and percentages are by weight.

EXAMPLE 1 fl-Carotene fermentation medium CM-l A series of 500 ml. flasks, each containing 100 ml. of fermentation medium CM-l or the medium disclosed in U.S. Pat. 3,079,380, Example 1, was prepared and inoculated using the procedure disclosed in Example 1 of U.S.

before sterilization).

1 Sonneborn and Sons T.M. for deodo iz d k closed in U.S. Pat. 3,025,221. r e erosene as dis TABLE I Medium disclosed in U.S. Pat. Fermentation 3,079,380, Example 1: medium CM-l Mg.-,6-carotene 912 1740 EXAMPLE 2 The conditions, inoculum, and medium examployed in U.S. Pat. 3,079,380, Example 2, were followed, except that the citrus molasses was varied to much higher levels. The results are shown in Table II.

1. In a method for microbiologically producing ,B-carotene in a submerged aerobic fermentation using Blakeslew trispora, the improvement which comprises incorporating at least about 7 /2% of citrus molasses in a B-carotene fermentation medium.

2. In a method for microbiologically producing B-carotene in a submerged aerobic fermentation using Blakeslea trispora, the improvement which comprises incorporating at least about 7 /3% of citrus molasses in a fi-carotene fermentation medium comprising non-hydrolyzed soybean meal, corn steep liquor, cotton seed oil, and deobase, and recovering the B-carotene so produced.

3. In a method for microbiologically producing fi-carotene in a submerged aerobic fermentation using Blakeslea trispora, the improvement which comprises the use of a fermentation medium consisting of the following ingredients:

Water, q.s. to 1 liter.

and adjusting the pH to 6.0 to 7.5 with NaOH before sterilization.

References Cited UNITED STATES PATENTS 3,013,948 12/1961 Hoffmann et a1. I-114 3,079,380 2/1963 Ciegler et al 19528 X 3,291,701 12/1966 Fulde 195-28 ALVIN E. TANENHOLTZ, Primary Examiner 

